KR100778942B1 - Fomitopsis pinicola Extract Having Anti-Inflammatory Effect and The Use Thereof - Google Patents

Abstract

The present invention relates to pine needles mushroom extract and its use. Since pine needles fruiting body hot water extract of the present invention has an inhibitory effect on edema, it can be widely used in the development of a composition for the prevention and treatment of various inflammation-related diseases.

Pine Butterfly Extract, Anti-inflammatory, Anti-inflammatory

Description

Pineapple Butterfly Extract Having Anti-Inflammatory Effect and Its Use {Fomitopsis pinicola Extract Having Anti-Inflammatory Effect and The Use Thereof}

The present invention relates to pine needles mushroom extract and its use. More specifically, the present invention relates to pine needles fruiting fruit hot water extract exhibiting an anti-inflammatory effect and its use.

Inflammatory reactions are associated with various mediators and immune cells of local blood vessels and body fluids when infected with tissues (cells) or external infectious agents (bacteria, fungi, viruses, and various types of allergens). It has a series of complex physiological reactions, including substance secretion, fluid infiltration, cell migration, and tissue destruction, as well as external symptoms such as erythema, edema, fever, and pain. In normal cases, the inflammatory response removes external infectious agents and regenerates damaged tissues to restore the function of life.However, if the inflammatory response is excessive or persistent due to the absence of antigens or internal substances, it promotes mucosal damage. Some results lead to diseases such as cancer.

In vivo, inflammation causes various biochemical phenomena, and in particular, enzymes related to biosynthesis of nitric oxide synthase (NOS) and prostaglandins, enzymes that generate nitric oxide (NO) They are known to play an important role in mediating the inflammatory response. Therefore, cyclooxygenase (COX), an enzyme involved in synthesizing prostaglandins from nitric oxide synthase or arachidonic acid, an enzyme that produces NO from L-arginine, It is a major goal in blocking inflammation.

According to recent research, there are several types of NOS, such as brain NOS in the brain, neuronal NOS in the nervous system, and endothelial NOS in the vascular system. It is expressed at a level, and a small amount of NO produced by them plays an important role in maintaining normal homeostasis, such as inducing neurotransmission and vasodilation, whereas iNOS induced by various cytokines or external stimulants NO, which is rapidly over-produced by induced NOS, is known to cause cytotoxicity and various inflammatory reactions, and chronic inflammation is associated with an increase in iNOS activity.

In addition, there are two kinds of isomers of cyclooxygenase. While cyclooxygenase-1 (COX-1) is always present in cells and exhibits the action of synthesizing prostaglandins (PGs) necessary for cytoprotective action, COX -2 is known to play an important role in causing the inflammatory response by rapidly increasing in the cell during the inflammatory response. Transcriptional inflammatory factors, including iNOS and COX-2, which enhance the degree of NO and PGs, are involved in the pathogenesis of chronic diseases including various sclerosis, Parkinson's disease, Alzheimer's disease and colon cancer.

NF-kB is a nuclear protein of the Rel gene family. In the cytoplasm, NF-kB binds to I-kB and is in an inactive form. However, NF-kB, TNF-α (tumor necrosis factor-α), and IL-1 ( After I-kB kinase is activated by various stimuli such as chemokines (lipokpolysaccharide, LPS) such as interleukin-1), I-kB is released through phosphorylation. NF-kB composed of p50 and p65 heterodimers is known to promote gene expression that, after being activated, migrates to the nucleus and induces an inflammatory response.

Multifunctional cytokines such as TNF-α are not only expressed in normal tissues, but are also increased in the course of lesions, and play an important role in skin inflammation, particularly during cancer promotion. TNF-α has been reported to be associated with inflammatory skin diseases in humans. Symptoms have been alleviated by treatment with TNF-α antibodies to various inflammatory diseases and allergic phenomena, and TNF-α also acts as an endogenous cancer promoter by activating neutrophils and increasing hydrogen peroxide production. Therefore, by inhibiting the expression of the cytokine TNF-α, which plays a pivotal role in the inflammatory stage closely related to the oncogenic stage, or inhibiting the production of PGE ₂ due to COX-2 inactivation, It is likely that the lesion process that accompanies the increase can be controlled. When treated with antibiotics under bacterial infection, the lipid polysaccharide (LPS) released from the extracellular membrane killing bacteria causes endotoxin shock and, when severely destroyed by this process, does not neutralize the continued production of LPS. do. Therefore, drug treatment may be beneficial by reducing the action of LPS. Reduction of inflammation due to the use of nonsteroidal anti-inflammatory drugs (NSAIDs) to date has often resulted in pain relief for a considerable period of time, and most of the non-narcotic analgesics (such as aspirin) are also anti- It has an inflammatory effect and has been used properly for the treatment of acute and chronic inflammatory diseases. However, due to side effects, it is difficult to use in the long term, and as a result, the development of new or improved therapeutics is essential and urgent because the severity of the side effects is too great for the present treatment. In connection with the problem of overcoming these side effects, efforts have recently been made to find the active ingredient in natural products that are used in the private sector.

Fomitopsis pinicola ( Fomitopsis pinicola ) is a native mushroom of Korea. It has no sack, and the stem is attached to the tree trunk in the shape of a shelf and forms a semi-circular shape. The diameter of the pod is 30cm in diameter and 15cm thick. The band is enclosed and the bottom is yellowish white with fine pores. It is known that it has been used for the prevention and healing of various lifestyle diseases since ancient times. Recently, the inventors of the present invention have been concerned with the prevention of diabetic complications and improvement of diabetes-induced hypercholesterolemia (Korean patent application No. 2006-1430), pancreatic cell damage inhibitory effect (Korean patent application No. 2006-1431), The inhibitory effect of active oxygen production due to diabetes (Korean Patent Application No. 2005-133990) has been elucidated, and the active protein polysaccharide is a combination of β-1,3-glucan and β-1,6-heterogalactomannan. It has been found that the protein is bound to β-1,3-glucano-β-1,6-heterogalactomannan having a molecular weight of 300,000 to 500,000.

However, in addition to the above effects of pine beetle mushroom extract is being researched to prove a variety of effects, as a natural product having an anti-inflammatory effect that can replace the non-steroidal anti-inflammatory drugs or non-narcotics analgesics used in the existing inflammation treatment Therefore, the necessity of research on this is emerging.

Thus, the present inventors, to determine the anti-inflammatory effect of pine berry butterfly extract, oral administration of pine berry butterfly extract, injecting carrageenan, an inflammation-causing substance, and the result of edema increase over time, the fact that it suppresses edema It was confirmed that the present invention was completed.

After all, the main object of the present invention is to provide an extract of pine fruit butterfly fruit fruit body exhibiting an anti-inflammatory effect.

It is another object of the present invention to provide a composition comprising an electric pine leaf butterfly fruiting hot water extract as a main, and exhibits an anti-inflammatory effect.

The present invention is a pine grass butterfly ( Fomitopsis) It provides a composition for the prevention and treatment of various inflammation-related diseases, including the pinicola ) extract as an active ingredient: wherein, pine berry butterfly extract is not particularly limited, but is a fruiting body hydrothermal extract, Inflammation-related diseases include, but are not limited to, rheumatoid arthritis, sepsis, asthma, acute pain, chronic pain, neuropathic pain, postoperative pain, pain such as joint pain, gouty arthritis, gout renal disease or inflammatory bowel disease. .

In addition, the present invention provides a health functional food comprising pine needles fruiting fruit hot water extract and food acceptable food supplement additives exhibiting a prophylactic effect of inflammation-related diseases: wherein the health functional food is not particularly limited thereto. However, it is preferable that the health drink.

The hot water extract of the fruiting body of the pine needles butterfly is obtained by cutting, crushing, watering and heating the pine fruit butterfly, and then filtering it with three layers of cheese cloth. In addition, after the extraction is concentrated and precipitated with ethanol, the precipitate is stored by the method of lyophilization. In this case, the valence is not particularly limited thereto, but it is preferable to add purified water corresponding to about 30 times the weight of the crushed fruiting body, and the extraction is not particularly limited thereto, but the extraction is preferably performed at 100 ° C. for about 24 hours. However, the concentration is not particularly limited, but it is preferable to concentrate at 40 ° C. until it is about 1/10 of the initial volume.

The anti-inflammatory effect of the hot-water extract of pine needles mushroom fruiting body of the present invention obtained from the above method was investigated as follows. After oral administration of three kinds of hot-water extracts of pine beetle mushrooms of different concentrations, the volume of the right hind limb of the rats that caused edema with carrageenan was measured. Edema inhibition was calculated. The carrageenan is a sulfated polygalactan obtainable from algae, and in particular, λ-carrageenan has a characteristic of causing acute inflammation by selective toxic action on macrophages, and also activates plasmin. It has the property of enhancing the production of kinin.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example

Example 1 : Preparation of fruiting body hot water extract of pine grass butterfly

150 g of two-year-old pine pinnacle mushroom fruiting bodies provided from regenerated agricultural products in Pohang were cut into 1 × 1 cm, crushed, and then, 4.0L of purified water was added and extracted for 24 hours at 100 ° C. using a hot water extractor. Filtration with a cloth gave a hydrothermal extract. Further, the extracted extract was concentrated at 40 ° C. until the extract became 400 mL, ethanol was added to precipitate, and the precipitated alcohol insoluble substance was lyophilized and stored.

Example 2 Anti-inflammatory Effect of Hot Water Extract of Pine Fruits

Example 2-1 : Experimental group and test method

SD male rats with a body weight of 180 to 190 g were reared for 2 weeks or more in an experimental animal breeding room. The kennel was controlled by artificial lighting to adjust the lighting time from 8 am to 8 pm and 12 hours, and the room temperature was maintained at 24 ± 2 ℃. Six rats of 200 ± 5g body weight were used as a group, and the volume of the right hind limb (marked at a constant part of the ankle) of each group of animals was measured immediately before the injection of carrageenan, an inflammation-causing substance, according to Niemegeer et al. , 50 μl of 1% carrageenan (Sigma Chem) dissolved in saline was injected into the midfoot of the right hind limb, and the right hind limb volume of the experimental animal was measured again after 30 minutes and 3 hours. 30 minutes before subcutaneous injection of carrageenan, pine needle butterfly mushroom group 1 (WE1: 100mg / kg), hydrothermal extract group 2 (WE2: 200mg / kg), hydrothermal extract group 3 (WE3: 400mg / kg) and control group Control: The same amount of saline was administered orally. The edema growth rate was calculated by the ratio (%) to the volume before and after drug administration as shown in Equation 1 below.

Equation 1 :

Edema growth rate (%) = {( _after hind paw _edema -hind paw _{edema development} ) / hind paw _edema }} 100

Example 2-2 anti-inflammatory effect

The edema growth rate of male rats carried out by the method of Example 2-1 was measured, and the effects of the hot-spring extract of pine needles on the carrageenan-induced edema growth rate were examined. Statistical processing of the measured values between each experimental group was used the student's t-test (see Table 1).

Table 1 : Effect of Pine-napbi Mushroom Hot Water Extract on Carrageenan-Induced Edema Growth Rate in Male Rats

Experimental group ^{1 )} Edema of hind paws (%) After 30 minutes (%) % After 3 hours  Control 10.5 ± 1.61 ²⁾ 63.5 ± 7.3  WE1 10.4 ± 1.1 51.3 ± 6.2  WE2 9.9 ± 1.5 39.2 ± 5.5 ^*  WE3 10.2 ± 1.8 42.7 ± 5.1 ^*

¹⁾ Control: physiological saline administration group, WE1: pine zanbi mushroom hot water extract (100mg / kg) oral administration group, WE2: pine zanbi mushroom hot water extract (200mg / kg) oral administration group, WE3: pine zanbi mushroom hot water extract (400mg / kg) Oral administration group

²⁾ Mean ± standard deviation ( ^* : p <0.05) for 6 groups

Based on the results in Table 1, the edema inhibition rate of each experimental group was calculated as in Equation 2 below. The edema inhibition rate was calculated as the ratio of the edema growth rate of the drug administration group to the edema growth rate of the control group. Edema inhibition is shown in Table 2 below.

Equation 2 :

Edema inhibition rate (%) = {(edema growth rate of control group-edema growth rate of each experimental administration group) / control edema growth rate} × 100

Table 2 : Anti-inflammatory Effects of Hot Water Extracts of Pine Needles on Carrageenan-Induced Inflammation in Male Rats

Experimental group ^{1 )} Edema inhibition rate (%) After 30 minutes (%) % After 3 hours  WE1 1.0 19.2  WE2 5.7 38.2  WE3 2.9 32.7

¹⁾ Same as Table 1

As shown in Table 2, when the pine needle butterfly hot water extract was administered, the increase in edema observed after 30 minutes of carrageenan injection did not show a significant difference between the control group and the pine needle butterfly hot water extract administration group, but the edema observed after 3 hours The increase rate was significantly decreased compared to the inflammatory control group. Especially, in the group of 200mg / kg and 400mg / kg pine needle butterfly extract, the inhibition rate was about 38.2% and 32.7%, respectively.

Edema caused by carrageenan, an inflammation-induced model drug, is usually induced by histamine or serotonin in phase 1 and in phase 2 induced by kinins, prostaglandins, and slow reactive substance-A (SRS-A). It is known that phase 1 edema occurs immediately after carrageenan injection and reaches a peak at about 30 minutes, and then disappears. Phase 2 edema occurs after 1 hour of carrageenan injection and is known to persist for a long time. As a result of the experiment, various pine zanbi mushroom extract did not have a significant effect on the first phase edema induced by histamine or serotonin, but it was confirmed that it has anti-inflammatory action that inhibits the second phase edema induced by prostaglandins and the like. It became.

Since pine needles fruiting body hot water extract of the present invention has an anti-inflammatory effect, it can be widely used in the development of a composition for the prevention and treatment of various inflammation-related diseases.

Claims (7)

A pineapple butterfly ( Fomitopsis pinicola ) comprising the extract as an active ingredient, and the composition for preventing and treating inflammation showing inhibitory activity against carrageenan-induced inflammation. The method of claim 1, The pine twig butterfly extract is characterized in that the fruiting body hydrothermal extract. The method of claim 2, The hydrothermal extract is extracted by cutting, crushing, watering and heating pine pinnacus mushroom fruiting body, and then obtaining the composition. The method of claim 3, wherein After extracting at 100 ° C. for 24 hours, the composition is characterized by filtration with three layers of cheese cloth. delete A functional food for preventing and improving inflammation, comprising pine needles, fruiting bodies, hot water extracts, and food acceptable additives, and showing inhibitory activity against carrageenan-induced inflammation. The method of claim 6, The health functional food is a health functional food, characterized in that the health drink.